The identification of new Plasmodium falciparum kinase targets have been hampered due to the limited numbers of kinases that have been purified and the lack of an available high-throughput assay to screen potential drug candidates. Our long-term goal is to develop a high-throughput biochemical screening platform to profile the selectivity of novel kinase inhibitors from companion parallel-synthetic libraries. The overall objective of this application is to develop both a model P. falciparum kinase-displaying T7 phage platform and a Bio-Layer Interferometry (BLI) assay amendable for high-throughput screening. The central hypothesis of this application is that P. falciparum kinase-displaying phage can be developed to screen putative anti-malarial P. falciparum kinase inhibitors. We plan to test our central hypothesis and accomplish the objective of this application by pursuing the following two specific aims: (1) Develop a phage display platform for P. falciparum kinases;and (2) Develop a biochemical assay amendable to high-throughput capable of direct determination of binding affinity of kinase inhibitors. The rationale for developing a BLI screening assay amendable for high- throughput is that in combination with parasitic proliferation assays it will allow for both the identification of essential malarial kinases and provide a screening platform for parallel-synthetic drug libraries. The expected outcome of the proposed work is that the necessary enabling technology will be developed for establishing a malarial kinase screening platform using phage and BLI. In combination with antimalarial assays it will then become possible to identify key malarial kinases as new drug targets. In addition, companion parallel-synthetic libraries of putative antimalarial kinase inhibitors can be later screened for specificity to particular P. falciparum kinases.
The overall objective of this application is to develop both a model Plasmodium falciparum kinase-displaying T7 phage platform and a Bio-Layer Interferometry (BLI) assay amendable for high-throughput screening. The expected outcome of the proposed work is that the necessary enabling technology will be developed for establishing a malarial kinase screening platform using phage and BLI. We anticipate this technology would allow antimalarial drug candidates from companion parallel synthetic kinase inhibitor libraries to be profiled for both affinity and specificity for relevant malarial kinase targets.
